Hiding on jagged karst pinnacles: A new microendemic genus and species of a limestone-dwelling agamid lizard (Squamata: Agamidae: Draconinae) from Khammouan Province, Laos.

We describe a unique new species and genus of agamid lizard from the karstic massifs of Khammouan Province, central Laos. Laodracon carsticola Gen. et sp. nov. is an elusive medium-sized lizard (maximum snout-vent length 101 mm) specifically adapted to life on limestone rocks and pinnacles. To assess the phylogenetic position of the new genus amongst other agamids, we generated DNA sequences from two mitochondrial gene fragments (16S rRNA and ND2) and three nuclear loci ( BDNF, RAG1 and c-mos), with a final alignment comprising 7 418 base pairs for 64 agamid species. Phylogenetic analyses unambiguously place the new genus in the mainland Asia subfamily Draconinae, where it forms a clade sister to the genus Diploderma from East Asia and the northern part of Southeast Asia. Morphologically, the new genus is distinguished from all other genera in Draconinae by possessing a notably swollen tail base with enlarged scales on its dorsal and ventral surfaces. Our work provides further evidence that limestone regions of Indochina represent unique "arks of biodiversity" and harbor numerous relict lineages. To date, Laodracon carsticola Gen. et sp. nov. is known from only two adult male specimens and its distribution seems to be restricted to a narrow limestone massif on the border of Khammouan and Bolikhamxai provinces of Laos. Additional studies are required to understand its life history, distribution, and conservation status.

A new species of Cyrtodactylus Gray, 1827 (Squamata: Gekkonidae) from the Thai-Malay Peninsula and the independent evolution of cave ecomorphology on opposite sides of the Gulf of Thailand.

An integrative taxonomic analysis recovers a distinctive new species of the gekkonid genus Cyrtodactylus Gray, 1827 from Satun Province in extreme southern Thailand as the sister species to the Cyrtodactylus intermedius group of southern Indochina, approximately 600 km to the northeast across the Gulf of Thailand. Based on 1449 base pairs of the mitochondrial gene NADH dehydrogenase subunit 2 (ND2) and its flanking tRNAs, the new species, C. disjunctus sp. nov., bears a pairwise sequence divergence from the mean divergences of the intermedius group species ranging from 17.923.6%. Three different principal component analyses (PCA) and a multiple factor analysis (MFA) recover C. disjunctus sp. nov. as a highly distinctive karst cave-adapted species based on morphology and color pattern. Its sister species relationship to the intermedius groupto which it is added herefurther underscores a growing body of analyses that have recovered a trans-Gulf of Thailand connection across the submerged Sunda Shelf between the southern Thai-Malay Peninsula and southern Indochina. Fragmented karstic archipelagos stretching across Indochina have served as foci for the independent evolution of nearly 25% of the species of Cyrtodactylus. The description of C. disjunctus sp. nov. continues to highlight the fact that karstic habitats support an ever-increasing number of threatened site-specific endemics that compose much of the reptile diversity of many Asian nations but, as of yet, most of these landscapes have no legal protection.

A new species of stream toad of the genus Ansonia Stoliczka, 1870 (Anura: Bufonidae) from Nakhon Si Thammarat Range in southern Thailand.

An integrative taxonomic analysis confirmed the new species status of a recently discovered upland population of Ansonia from Thongsong, Thongsong District, Nakhon Si Thammarat Province, Thailand from the sky island archipelago south of the Isthmus of Kra. Ansonia infernalis sp. nov. is a member of the Thai-Burmese clade within the more inclusive Thai-Malaya Peninsula clade of western and southern Thailand. It is separated from all other species of Ansonia by a unique combination of morphometric and discrete morphological and color pattern characteristics and is the sister species of a clade of eight other species found north of the Isthmus of Kra. Ansonia infernalis sp. nov. is the newest member of a long list of range-restricted endemics from the sky island archipelago of the Thai-Malay Peninsula and continues to underscore the unexplored nature of this region and its need for conservation.

An integrative taxonomic revision of slug-eating snakes (Squamata: Pareidae: Pareineae) reveals unprecedented diversity in Indochina.

Slug-eating snakes of the subfamily Pareinae are an insufficiently studied group of snakes specialized in feeding on terrestrial mollusks. Currently Pareinae encompass three genera with 34 species distributed across the Oriental biogeographic region. Despite the recent significant progress in understanding of Pareinae diversity, the subfamily remains taxonomically challenging. Here we present an updated phylogeny of the subfamily with a comprehensive taxon sampling including 30 currently recognized Pareinae species and several previously unknown candidate species and lineages. Phylogenetic analyses of mtDNA and nuDNA data supported the monophyly of the three genera Asthenodipsas, Aplopeltura, and Pareas. Within both Asthenodipsas and Pareas our analyses recovered deep differentiation with each genus being represented by two morphologically diagnosable clades, which we treat as subgenera. We further apply an integrative taxonomic approach, including analyses of molecular and morphological data, along with examination of available type materials, to address the longstanding taxonomic questions of the subgenus Pareas, and reveal the high level of hidden diversity of these snakes in Indochina. We restrict the distribution of P. carinatus to southern Southeast Asia, and recognize two subspecies within it, including one new subspecies proposed for the populations from Thailand and Myanmar. We further revalidate P. berdmorei, synonymize P. menglaensis with P. berdmorei, and recognize three subspecies within this taxon, including the new subspecies erected for the populations from Laos and Vietnam. Furthermore, we describe two new species of Pareas from Vietnam: one belonging to the P. carinatus group from southern Vietnam, and a new member of the P. nuchalis group from the central Vietnam. We provide new data on P. temporalis, and report on a significant range extension for P. nuchalis. Our phylogeny, along with molecular clock and ancestral area analyses, reveal a complex diversification pattern of Pareinae involving a high degree of sympatry of widespread and endemic species. Our analyses support the "upstream" colonization hypothesis and, thus, the Pareinae appears to have originated in Sundaland during the middle Eocene and then colonized mainland Asia in early Oligocene. Sundaland and Eastern Indochina appear to have played the key roles as the centers of Pareinae diversification. Our results reveal that both vicariance and dispersal are responsible for current distribution patterns of Pareinae, with tectonic movements, orogeny and paleoclimatic shifts being the probable drivers of diversification. Our study brings the total number of Pareidae species to 41 and further highlights the importance of comprehensive taxonomic revisions not only for the better understanding of biodiversity and its evolution, but also for the elaboration of adequate conservation actions.

Confirmation of Leptobrachellaventripunctata (Fei, Ye, and Li, 1990), based on molecular and morphological evidence in Thailand.

BACKGROUND: Thailand is considered a global biodiversity hotspot that is known to harbour a striking diversity of endemic species. However, several research studies have determined that the level of amphibian diversity in the country has been significantly underestimated. The megophryid genus Leptobrachella Smith, 1925 is currently known to include 89 species that are primarily distributed throughout southern China and Southeast Asia; however, only seven species have been found in Thailand. NEW INFORMATION: Based on an integrative approach encompassing genetic and morphological analyses, we have concluded that the population identified from Chiang Rai Province of Thailand is conspecific with Leptobrachellaventripunctata (Fei, Ye, and Li, 1990). Importantly, this is the first confirmation record of this species, based on molecular and morphological evidence in Thailand. The discovery of this species reaffirms that the diversity within the genus has been underestimated with many species yet to be discovered. In addition, the findings of our study further highlight the lack of existing knowledge on amphibian taxonomy and an underestimation of the biodiversity that exists along these national border areas.

Integrative taxonomy reveals a new species of Leptobrachella (Anura: Megophryidae) from the mountains of northern Thailand.

The genus Leptobrachella is one of the most speciose and taxonomically troubling groups of Asian anurans. Herein, we describe a new species of Leptobrachella from Doi Inthanon, Chiang Mai Province, Thailand based on the integration of morphological, acoustic and molecular data. The new species, Leptobrachella murphyi sp. nov., is morphologically distinguished from its congeners on the basis of body size, dorsal skin texture and coloration, ventral coloration and pattern, degree of webbing and fringing on the fingers and toes, iris coloration in life, pattern of markings on flanks and pattern of body macroglands. It also differs from its phylogenetically close congeners by an uncorrected p-distance of >9.6% for a fragment of 16S rRNA. The advertisement call of the new species consists of 4.54.7 kHz (at 15 C) and without a distinct introductory note. Leptobrachella murphyi sp. nov. likely occurs across the Thanon Thong Chai Range and analyses provide evidence of unknown biodiversity and species composition on Doi (mountain) Inthanon. In addition, the congeneric species L. minima was also confirmed in Doi Inthanon. The coexistence pattern of Leptobrachella in Doi Inthanon deserves further study. As Thailands highest mountain and biodiversity reservoir, the need for further biological exploration is urgent given ongoing habitat loss and degradation.

First national record of Quasipaaverrucospinosa (Bourret, 1937) (Amphibia: Anura: Dicroglossidae) from Thailand with further comment on its taxonomic status.

BACKGROUND: Spiny Frog Quasipaa is a genus of frogs that belongs to a relatively poorly known group. Most of the species distribution has been recorded in China; however, a few incidences of identification have occurred in the eastern part of Indochina. To date, only one species (Quasipaafasciculispina) of Quasipaa has been recorded from Chanthaburi and Trat Provinces in south-eastern Thailand. NEW INFORMATION: Based on recent fieldwork conducted in northern Thailand, we report a new record of Quasipaaverrucospinosa from Doi Phu Kha National Park, Nan Province at an altitude of 900-1000 m a.s.l. Our study has demonstrated that populations of this species are paraphyletic and has revealed deep genetic differences. Therefore, it is recommended that a comprehensive study be undertaken to clarify the taxonomic and geographic distribution of this species for its suitable protection and conservation.

First records of the fanged frogs Limnonectes bannaensis Ye, Fei & Jiang, 2007 and L. utara Matsui, Belabut & Ahmad, 2014 (Amphibia: Anura: Dicroglossidae) in Thailand.

BACKGROUND: The taxonomic status of the Thai populations belonging to the Limnonectes kuhlii species complex is controversial, due to phenotypic similarity in the cryptic species complex. Recently, some studies on this group in Thailand have discovered four new species: L. taylori, L. megastomias, L. jarujini and L. isanensis. Even so, the diversity of this group is still incomplete. NEW INFORMATION: Based on an integrative approach encompassing genetic and morphological analyses, we conclude that the Limnonectes populations from Nan Province (northern) and Yala Province (southern) of Thailand are conspecific with L. bannaensis Ye, Fei & Jiang, 2007 and L. utara Matsui, Belabut & Ahmad, 2014, respectively. These are the first records of these species in Thailand. Our study highlights the importance of using DNA sequence data in combination with morphological data to accurately document species identity and diversity. This is especially important for morphologically cryptic species complexes and sympatrically occurring congeners.

First national record of Gracixalus quangi Rowley, Dau, Nguyen, Cao & Nguyen, 2011 and G. yunnanensis Yu, Li, Wang, Rao, Wu &Yang, 2019 (Amphibia: Anura: Rhacophoridae) from Thailand.

BACKGROUND: The bushfrog genus Gracixalus Delorme, Dubois, Grosjean & Ohler, 2005 is found in southern and south-western China, Vietnam, Laos, Thailand and Myanmar. It is presently comprised of 17 species. In Thailand, only two species have been recorded, namely G. carinensis (Boulenger) and G. seesom (Massui, Khonsue, Panha & Eto). The latter of these two species is currently known to be endemic to the country. NEW INFORMATION: Based on recent field work conducted in 2019 in Doi Phu Kha National Park, Nan Province of northern Thailand, we are reporting two new records of the genus Gracixalus, G. quangi and G. yunnanensis, from Thailand, based on morphological and molecular evidence. In addition, this is the first study to report on the identification of a female specimen of G. yunnanensis. Furthermore, morphological data and natural history notes of the aforementioned species in Thailand have been provided, along with updated locations for the distribution of both species.

A new karst-dwelling, colorful pitviper (Viperidae: Trimeresurus) from northern Peninsular Thailand.

We describe a colorful and distinctively patterned, karst-dwelling pitviper, Trimeresurus kuiburi sp. nov., from the isolated, coastal massif of Khao Sam Roi Yot in Kui Buri District, Prachuap Khiri Khan Province, in northern Peninsular Thailand. The new species, member of the 'Cryptelytrops group' sensu Malhotra Thorpe (2004) and morphologically and genetically allied to Trimeresurus kanburiensis and T. venustus, differs from all pitviper taxa by a combination of red/purple bands on a green dorsum; a white concave suborbital stripe in males (straight and less visible in females); white, spaced vertebral dots in males (absent in females); pale green belly lacking dark dots or stripe on the lateral sides of the ventrals; partially fused first supralabial and nasal scale; 19 dorsal scale rows at midbody; 164171 ventrals; 6365 subcaudals in males, 5153 in females; maximal known SVL of 451 mm; and long, papillose hemipenes.

Phylogenetic analyses of distantly related clades of bent-toed geckos (genus Cyrtodactylus) reveal an unprecedented amount of cryptic diversity in northern and western Thailand.

Cyrtodactylus species are the most diverse of the geckos and are widely distributed in Southeast Asia, including Thailand. However, their patterns of distribution, especially in northern and western parts of Thailand, remain unknown because few Cyrtodactylus species in these regions have been described. Thus, a data set of mitochondrial NADH dehydrogenase 2 (ND2) gene and flanking tRNAs from Cyrtodactylus found in northern and western Thailand, including contiguous areas, was assembled to elucidate phylogenetic relationships and identify the distribution patterns of these geckos. The results showed four well-supported clades, a northwestern clade (A), a northern clade (B), a western clade (C), and a special clade characterized by specific morphological features (D). Clades A-C were grouped with strong support by the geography of their localities from northern Thailand (Mae Hong Son and Chiang Mai Provinces) along the Tenasserim mountain ranges to Phang-Nga Province, Thailand. Clade D is a distinct clade of Cyrtodactylus species characterized by a tuberculate and prehensile tail and distributed widely in mainland Southeast Asia. Overall, the results suggest a pattern of geographic separation and distribution of Cyrtodactylus in northern and western Thailand. Additionally, this study provides evidence of a hidden biodiversity of Cyrtodactylus in these regions.

A new species of Hemiphyllodactylus Bleeker (Squamata; Gekkonidae) from Peninsular Thailand that converges in morphology and color pattern on Pseudogekko smaragdinus (Taylor) from the Philippines.

An integrative taxonomic analysis of newly discovered populations of Hemiphyllodactylus from Indochina recovered a new species from Peninsular Thailand and two others from Laos. The new Thai species, Hemiphyllodactylus pardalis sp. nov. described herein, is the sister species to all other species in the newly designated Indochina clade and has a unique spotted contrasting dorsal pattern not seen in any other species of Hemiphyllodactylus. This, along with its elongate and gracile body morphology, renders it remarkably convergent on the Philippine gecko Pseudogekko smaragdinus despite the fact it is deeply nested within Hemiphyllodactylus. The description of this new species underscores the depauperate nature of Hemiphyllodactylus diversity throughout Indochina in general and Thailand in particular compared to neighboring upland areas of Myanmar and Peninsular Malaysia. This especially underscores the need for continued herpetofaunal field surveys in montane areas of western Thailand.

A new species of Cnemaspis Strauch (Squamata: Gekkonidae) of the C. siamensis group from Tenasserim Mountains, Thailand.

An integrative taxonomic analysis recovered the new species Cnemaspis selenolagus sp. nov. of the C. siamensis group as the sister species to C. punctatonuchalis. The new species was discovered in mountain evergreen tropical forests of in Suan Phueng District of Ratchaburi Province, western Thailand. Additionally, the analysis recovered a deep genetic divergence between northern and southern clades within the C. siamensis group that occur on opposite sides of the Isthmus of Kra-a well-known biogeographic region of cladogenic turnover. The description of C. selenolagus sp. nov. brings the total number of species of Cnemaspis in Thailand to 16, 11 of which compose the C. siamensis group-a lineage endemic to the Thai-Malay Peninsula. This underscores the physiographic complexity of this narrow peninsula in that it can support a large number of closely related species in only the northern two-thirds of its length.

A new species Cyrtodactytlus Gray (Squamata: Gekkonidae) from western Thailand and the phylogenetic placement of C. inthanon and C. doisuthep.

A new species of Cyrtodactylus from Tak Province, Thailand, Cyrtodactylus amphipetraeus sp. nov., is described using an integrative taxonomic analysis based on morphology, color pattern, and the mitochondrial gene NADH dehydrogenase subunit 2 (ND2). The phylogenetic analyses place the new species within the C. sinyineensis group which was previously thought to be endemic to the Salween Basin in southern Myanmar. The phylogeny also places C. inthanon in the C. sinyneensis group which is expanded herein to also include the group's sister species C. doisuthep. Along with C. amphipetraeus sp. nov., these are the first three species of the C. sinyineensis group to be found outside of Myanmar east of the Tenasserim Mountains. The Tenasserim Mountain region is discussed as an area of cladogeneic turnover.

A new species of Micryletta (Amphibia: Microhylidae) from southern Thailand.

We report on a new species, Micryletta dissimulans sp. nov., from the lowland forests of southern Thailand, which is described based on molecular and morphological evidence. The new species is characterized by a combination of the following characters: small body size (20.3-22.4 mm in males, 24.4-26.7 mm in females); slender body habitus; head longer than wide; snout rounded in dorsal and lateral view; eye length equal to snout length; tibiotarsal articulation reaching to tympanum; dorsal surface slightly granulated to shagreened; supratympanic fold indistinct, ventrally edged in black with large black spot behind eye; outer metatarsal tubercle absent; dorsum reddish-brown with merging irregular-shaped brown blotches edged in beige, no black spots on dorsum; body flanks brown with large black spots edged in whitish mottling, two large black blotches in axillary and inguinal areas on each side; lateral sides of head black, with white patches on lips absent, whitish mottling on tympanum and axillary region; ventral surface pinkish to bluish-gray, translucent, laterally with dark-brown marbled pattern, medially immaculate; throat in males dark-gray with sparse white mottling laterally; iris copper-orange. The new species is divergent from all other congeners in 16S rRNA gene sequences (5.0%-7.4%). To date, Micryletta dissimulans sp. nov. is only known from a single locality in Saba Yoi District, Songkhla Province, Thailand, at an elevation of 120 m a.s.l., but is also expected to occur in neighboring parts of Malaysia. We suggest Micryletta dissimulans sp. nov. be considered as a Data Deficient (DD) species following the IUCN's Red List categories (IUCN Standards and Petitions Committee, 2019).

A little frog leaps a long way: compounded colonizations of the Indian Subcontinent discovered in the tiny Oriental frog genus Microhyla (Amphibia: Microhylidae).

Frogs of the genus Microhyla include some of the world's smallest amphibians and represent the largest radiation of Asian microhylids, currently encompassing 50 species, distributed across the Oriental biogeographic region. The genus Microhyla remains one of the taxonomically most challenging groups of Asian frogs and was found to be paraphyletic with respect to large-sized fossorial Glyphoglossus. In this study we present a time-calibrated phylogeny for frogs in the genus Microhyla, and discuss taxonomy, historical biogeography, and morphological evolution of these frogs. Our updated phylogeny of the genus with nearly complete taxon sampling includes 48 nominal Microhyla species and several undescribed candidate species. Phylogenetic analyses of 3,207 bp of combined mtDNA and nuDNA data recovered three well-supported groups: the Glyphoglossus clade, Southeast Asian Microhyla II clade (includes M. annectens species group), and a diverse Microhyla I clade including all other species. Within the largest major clade of Microhyla are seven well-supported subclades that we identify as the M. achatina, M. fissipes, M. berdmorei, M. superciliaris, M. ornata, M. butleri, and M. palmipes species groups. The phylogenetic position of 12 poorly known Microhyla species is clarified for the first time. These phylogenetic results, along with molecular clock and ancestral area analyses, show the Microhyla-Glyphoglossus assemblage to have originated in Southeast Asia in the middle Eocene just after the first hypothesized land connections between the Indian Plate and the Asian mainland. While Glyphoglossus and Microhyla II remained within their ancestral ranges, Microhyla I expanded its distribution generally east to west, colonizing and diversifying through the Cenozoic. The Indian Subcontinent was colonized by members of five Microhyla species groups independently, starting with the end Oligocene-early Miocene that coincides with an onset of seasonally dry climates in South Asia. Body size evolution modeling suggests that four groups of Microhyla have independently achieved extreme miniaturization with adult body size below 15 mm. Three of the five smallest Microhyla species are obligate phytotelm-breeders and we argue that their peculiar reproductive biology may be a factor involved in miniaturization. Body size increases in Microhyla-Glyphoglossus seem to be associated with a burrowing adaptation to seasonally dry habitats. Species delimitation analyses suggest a vast underestimation of species richness and diversity in Microhyla and reveal 15-33 undescribed species. We revalidate M. nepenthicola, synonymize M. pulverata with M. marmorata, and provide insights on taxonomic statuses of a number of poorly known species. Further integrative studies, combining evidence from phylogeny, morphology, advertisement calls, and behavior will result in a better systematic understanding of this morphologically cryptic radiation of Asian frogs.

A new species of the genus Xenophrys Anura Megophryidae from northern Thailand.

Species of Xenophrys are conserved morphologically and live primarily in forests. In Thailand, the genus harbors many cryptic species. Herein we report the collection of specimens from Doi Inthanon, Chiang Mai Province, northern Thailand, which were identified previously as X. minor. Molecular and morphological analyses find that these specimens differ significantly from other known congeners, and therefore we describe a new species. Further, our phylogenetic analyses indicate that X. latidactyla is a junior synonym of X. palpebralespinosa.

On the road to Mandalay: contribution to the Microhyla Tschudi, 1838 (Amphibia: Anura: Microhylidae) fauna of Myanmar with description of two new species.

We present a morphological and molecular assessment of the Microhyla fauna of Myanmar based on new collections from central (Magway Division) and northern (Kachin State) parts of the country. In total, six species of Microhyla are documented, including M. berdmorei, M. heymonsi, M. butleri, M. mukhlesuri and two new species described from the semi-arid savanna-like plains of the middle part of the Irrawaddy (Ayeyarwady) River Valley. We used a 2 481 bp long 12S rRNA-16S rRNA fragment of mtDNA to hypothesize genealogical relationships within Microhyla. We applied an integrative taxonomic approach combining molecular, morphological, and acoustic lines of evidence to evaluate the taxonomic status of Myanmar Microhyla. We demonstrated that the newly discovered populations of Microhyla sp. from the Magway Division represent two yet undescribed species. These two new sympatric species are assigned to the M. achatina species group, with both adapted to the seasonally dry environments of the Irrawaddy Valley. Microhyla fodiens sp. nov. is a stout-bodied species with a remarkably enlarged shovel-like outer metatarsal tubercle used for burrowing and is highly divergent from other known congeners (P-distance≥8.8%). Microhyla irrawaddy sp. nov. is a small-bodied slender frog reconstructed as a sister species to M. kodial from southern India (P-distance=5.3%); however, it clearly differs from the latter both in external morphology and advertisement call parameters. Microhyla mukhlesuri is reported from Myanmar for the first time. We further discuss the morphological diagnostics and biogeography of Microhyla species recorded in Myanmar.